Isolation and characterization of the aureobasidin A-resistant gene, aur1R, on Schizosaccharomyces pombe: roles of Aur1p+ in cell morphogenesis.
To study the mechanism of action of the antibiotic aureobasidin A (AbA) on yeasts, we isolated a dominant mutant of Schizosaccharomyces pombe which gave high resistance to AbA. From a genomic library of the mutant, an aur1R mutant gene conferring AbA resistance was isolated. One amino-acid mutation, a substitution of glycine with cysteine at residue 240, was responsible for the acquisition of AbA resistance. The wild-type aur1+ gene was essential for viability, and its over-expression enhanced significant resistance to AbA. The predicted protein of S. pombe aur1R was highly homologous in primary structure and hydropathy profile with that of Saccharomyces cerevisiae AUR1R isolated as an AbA-resistance gene. To analyze a role in cell growth of S. pombe aur1+, temperature-sensitive mutants (aur1ts) were obtained by random mutagenesis procedures using a modified PCR. The aur1ts mutation caused a defect in cell elongation at the non-permissive temperature and finally led to cell death. These results suggest that Aur1p was a target of the antibiotic AbA and was required in the cell elongation of cell-end tips and in the viability of S. pombe.
Hashida-Okado
T
T
Biotechnology Research Laboratories, Takara Shuzo Co. Ltd., Seta 3-4-1, Otsu, Shiga, 520-21, Japan. okadot@takara.co.jp
Yasumoto
R
R
Endo
M
M
Takesako
K
K
Kato
I
I
D000595:Amino Acid Sequence; D000935:Antifungal Agents / Q000494:pharmacology*; D001483:Base Sequence; D048429:Cell Size / Q000187:drug effects / Q000235:genetics; D003001:Cloning, Molecular; D047630:Depsipeptides; D004352:Drug Resistance, Microbial; D005656:Fungal Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000502:physiology*; D015966:Gene Expression Regulation, Fungal; D006602:Hexosyltransferases; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D010456:Peptides, Cyclic / Q000494:pharmacology; D012568:Schizosaccharomyces / Q000166:cytology* / Q000235:genetics* / Q000254:growth & development
D000935:Antifungal Agents; D047630:Depsipeptides; D005656:Fungal Proteins; D010456:Peptides, Cyclic; C071398:aureobasidin A; D006602:Hexosyltransferases; C088714:phosphatidylinositol-ceramide phosphoinositol transferase